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Beijerinckia derxii releases plant growth regulators and amino acids in synthetic media independent of nitrogenase activity.

AIMS: This study aims at evaluating the ability of Beijerinckia derxii, a free-living nitrogen (N)-fixing bacterium frequently isolated from tropical soils, to release certain plant growth regulators [indoleacetic acid (IAA), ethylene, polyamines] and amino acids into the growth medium. METHODS AND RESULTS: The production of those substances was compared using both cultures in which nitrogenase was active (N-free medium) and cultures in which nitrogenase was repressed (combined-N cultures). Those cultures were grown under agitation and in absence of agitation. Total IAA production was higher in agitated, N-free cultures but specific production was greater in combined-N cultures under agitation. Putrescine and spermidine were detected under all conditions tested. Ethylene was produced in both N-free and combined-N cultures. A greatest diversity of amino acids was released in N-free cultures. CONCLUSIONS: There was no inhibition of the production of the analysed substances under conditions where nitrogenase was inactive. SIGNIFICANCE AND IMPACT OF THE STUDY: Beijerinckia derxii is potentially a producer of plant-active substances; its presence in the natural environment suggests that this bacterium may contribute to the development of other living organisms.

Amino Acids↗

Determination of urinary indolic metabolites.

A rapid mass spectral assay for tryptophol, 5-hydroxytryptophol, and 3-indoleacetic acid, employing stable-isotope labeled internal standards, is described. The compounds were extracted from urine or buffer with ethyl acetate and quantitatively measured by chemical-ionization mass spectrometry. The calibration curves were linear over a range of 0.06--2.8 microgram/ml. The technique was applied to the analysis of urine from patients with carcinoid tumors. In addition, the first synthesis of 5-methoxy-3-indoleacrylic acid is reported.

Carcinoid Tumor↗

Defined conditions for the initiation and growth of cotton callus in vitro. I. Gossypium arboreum.

Defined in vitro conditions for callus initiation by Gossypium arboreum L. were determined, and different tissues were evaluated as explant sources, Environmental conditions tested included light versus dark, and low light versus high light. Different nutrient media as well as carbohydrate sources were examined. Our data show that hypocotyl tissue was superior to cotyledon or leaf tissue as the explant source for callus proliferation; the Murashige-Skoog inorganic formulation with (in mg per 1) 100 myo-inositol, 0.4 thiamine-HCl, 2 indoleacetic acid (IAA), 1 kinetin, and 3% glucose solidifield by agar was the best medium to initiate callus. Cultures with sucrose as a carbohydrate source browned rapidly. Callus proliferation was superior under high light (8000 to 9000 lux) conditions at 20 +/- 1 degree C. Various combinations of auxins and cytokinins were tested for their ability to improve callus proliferation and subsequent growth of subcultures. Although the MS medium containing IAA and kinetin was found superior for obtaining rapid proliferation of callus from hypocotyl explants, a second medium containing 2 mg per 1 naphthaleneacetic acid (NAA) and 0.5 to 1 mg per 1 benzyladenine (BA) was found necessary for vigorous growth of subcultured callus. A MS medium with 5 to 10 mg per 1 N6-[delta2-isopentenyl]-adenine (2ip) and 1 mg per 1 NAA was also favorable for continued subculturing.

Ascorbic Acid↗

Plant growth regulators and amino acids released by Azospirillum sp in chemically defined media.

AIMS: To investigate the ability of Azospirillum sp., a facultative endophitic diazotrophic bacterium, to release plant growth regulators (PGR) such as polyamines, ethylene, indoleacetic acid and amino acids in both combined-N and N-free cultures. METHODS AND RESULTS: The presence of those substances was analysed by HPLC. Azospirillum sp. is capable of releasing PGR and amino acids into the culture medium. CONCLUSIONS: The type and quantity of the released substances varied, depending on the presence of combined-N in the medium. SIGNIFICANCE AND IMPACT OF THE STUDY: A better knowledge of PGR produced by Azospirillum sp. has been gained.

Amino Acids↗

Inhibition of auxin movement from the shoot into the root inhibits lateral root development in Arabidopsis.

In roots two distinct polar movements of auxin have been reported that may control different developmental and growth events. To test the hypothesis that auxin derived from the shoot and transported toward the root controls lateral root development, the two polarities of auxin transport were uncoupled in Arabidopsis. Local application of the auxin-transport inhibitor naphthylphthalamic acid (NPA) at the root-shoot junction decreased the number and density of lateral roots and reduced the free indoleacetic acid (IAA) levels in the root and [3H]IAA transport into the root. Application of NPA to the basal half of or at several positions along the root only reduced lateral root density in regions that were in contact with NPA or in regions apical to the site of application. Lateral root development was restored by application of IAA apical to NPA application. Lateral root development in Arabidopsis roots was also inhibited by excision of the shoot or dark growth and this inhibition was reversible by IAA. Together, these results are consistent with auxin transport from the shoot into the root controlling lateral root development.

Arabidopsis↗

Protein-protein interactions among the Aux/IAA proteins.

The plant hormone indoleacetic acid (IAA) transcriptionally activates early genes in plants. The Aux/IAA family of early genes encodes proteins that are short-lived and nuclear-localized. They also contain a putative prokaryotic betaalphaalpha DNA binding motif whose formation requires protein dimerization. Here, we show that the pea PS-IAA4 and Arabidopsis IAA1 and IAA2 proteins perform homo- and heterotypic interactions in yeast using the two-hybrid system. Gel-filtration chromatography and chemical cross-linking experiments demonstrate that the PS-IAA4 and IAA1 proteins interact to form homodimers in vitro. Deletion analysis of PS-IAA4 indicates that the betaalphaalpha containing acidic C terminus of the protein is necessary for homotypic interactions in the yeast two-hybrid system. Screening an Arabidopsis lambda-ACT cDNA library using IAA1 as a bait reveals heterotypic interactions of IAA1 with known and newly discovered members of the Arabidopsis Aux/IAA gene family. The new member IAA24 has similarity to ARF1, a transcription factor that binds to an auxin response element. Combinatorial interactions among the various members of the Aux/IAA gene family may regulate a variety of late genes as well as serve as autoregulators of early auxin-regulated gene expression. These interactions provide a molecular basis for the developmental and tissue-specific manner of auxin action.

Amino Acid Sequence↗

A rapid and sensitive auxin binding system for detecting N6-substituted adenines, and some urea and thiourea derivatives, that show cytokinin activity in cell division tests.

A selective, sensitive and rapid (2 min or less) method for detecting compounds with potential for cytokinin activity is described. The method does not measure cytokinesis; instead, it determines the ability of cytokinin-active agents to (i) activate the intake of either L-tryptophan or indoleacetic acid by germinated spores of the water-mould Achlya, while inhibiting the energy-dependent transport of all L-amino acids usually found in proteins; (ii) inhibit the energy-dependent transport of nucleosides and sugars by the same organism. The compounds with cytokinin activity generally activate auxin (tryptophan) intake at 10(-8) M or greater and inhibit at 10(-6) M or greater. The most effective activating compounds were N6-(delta2-isopentenyl)adenine, N6-benzyladenine. N6-furfuryladenine, and N6-(trans-hydroxy-3-methyl-but-2-enyl)adenine. These compounds are classed generally as cytokinins in plant growth studies. A cell membrane - localized glycopeptide of molecular weight 6000 was isolated from this organism and shown to be the site at which cytokinins, auxin, and tryptophan bind. An earlier study had also established that calcium ions bind to this entity as well. Tryptophan binding to the glycopeptide was enhanced by cytokinins, suggesting that this may be the way in which whole cells display enhanced tryptophan binding in the bioassay. On the other hand, calcium binding was antagonized by cytokinin. The results suggest that this may be an important experimental system for use in studying one possible way in which cytokinin may regulate plant growth.

Adenine↗

Reserpine augmentation of desipramine in refractory depression: clinical and neurobiological effects.

Early studies showed dramatic improvement in some depressed patients when a brief course of parenteral reserpine was added to ineffective tricyclic antidepressant (TCA) treatment. We treated eight patients with DSM-III melancholic major depression with desipramine (DMI) greater than or equal to 2.5 mg/kg/day (plasma levels greater than 125 ng/ml) for at least 4 weeks. All patients failed to respond and received reserpine 5 mg IM b.i.d. over 2 days, in seven cases as a placebo-controlled, double-blind trial. One patient had dramatic resolution of depressive and psychotic symptoms within 48 h, but relapsed within 2 weeks; two other patients had transient hypomanic symptoms. Depression ratings did not significantly change for the sample as a whole, but plasma and cerebrospinal fluid (CSF) levels of 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG) decreased and CSF levels of homovanillic acid (HVA) and 5-hydroxy-indoleacetic acid (5-HIAA) increased. Despite robust effects on central monoamine metabolism, reserpine augmentation appears insufficiently effective for routine use in managing refractory depression.

Adult↗

Chlorpromazine-induced alterations in hypothalamic amine metabolism and stress responses in severe cold.

To investigate the effects on the central nervous system of severe cold stress with and without chlorpromazine, guinea pigs were treated with chlorpromazine or 0.9% NaCl and exposed to -20 degrees C or +23 degrees C for 1 h. Hypothalamic noradrenaline (NA), dopamine (DA), 5-hydroxy-tryptamine (5-HT), 3-methoxy-4-hydroxyphenyl ethylene glycol (MHPG), homovanillinic acid (HVA) and 5-hydroxy-indoleacetic acid (5-HIAA) were determined by high-performance liquid chromatography. Serum, urinary and vitreous fluid catecholamines, muscle and liver glycogen, and blood glucose were also measured. Chlorpromazine caused distinct hypothermia at -20 degrees C and slight hypothermia at +23 degrees C. The rise in hypothalamic MHPG, 5-HIAA and MHPG/NA and in 5-HIAA/5-HT ratios in the cold indicate increased noradrenergic and serotonergic activity. The latter was inhibited by chlorpromazine and a drug-induced inhibition of noradrenergic neurons could not be ruled out. Chlorpromazine increased the turnover of DA at room temperature and the same tendency was seen in the cold. The hypothermic animals had low serum catecholamines, indicating diminished sympathetic activity. The chlorpromazine-treated cold-exposed animals did not react to the environmental stress by sympathetic activation, as urinary NA and adrenaline were not elevated, but DA was excreted by all the drug-treated animals. Vitreous fluid NA and DA were elevated as an indicator of cold stress, and no drug effect was seen in this fluid.

Animals↗

In vivo evaluation by differential pulse voltammetry of the effect of thyrotropin-releasing hormone (TRH) on dopaminergic and serotoninergic synaptic activity in the striatum and nucleus accumbens of the rat.

In vivo differential pulse voltammetry was used to determine the effect of thyrotropin-releasing hormone (TRH) on dopaminergic and serotoninergic synaptic activity in the striatum and nucleus accumbens of the rat. Thyrotropin releasing hormone (TRH) produces marked stimulatory effects on behaviour, which have been attributed to the release of dopamine in the nucleus accumbens. Other studies indicate a close relationship between the peptide and serotonin in the brain. We have thus used an improved differential pulse voltammetry technique to evaluate the effects of TRH on the extracellular content of the dopamine and serotonin metabolites. Dihydroxyphenylacetic acid (DOPAC) and 5-Hydroxy-indoleacetic acid (5HIAA) in the nucleus accumbens and striatum of the rat in vivo. TRH rapidly increased extracellular DOPAC, reaching a maximum after 60 min in the nucleus accumbens, and after 40 min in the striatum. There was also a slower increase in extracellular 5-HIAA content in both areas, reaching a plateau after 100 min. The delayed time course of the increase in 5-HIAA suggested that the increase in 5-HIAA content might be secondary to the increase in dopamine turnover produced by TRH. These results suggest that doses of TRH which produce behavioural stimulation increase the release of both dopamine and serotonin in the nucleus accumbens and striatum.

3,4-Dihydroxyphenylacetic Acid↗